Figure 2.

TNFα was the main mediator released by engaged CAR-T cells in inducing endothelial pro-inflammatory response. (A) The cytokine in the co-cultured supernatant of CAR-T/Nalm6 (sCAR-T) was analyzed by Luminex. (B) HUVEC were stimulated with TNFα (10 μg/ml), IFNγ (50 μg/ml), IL8 (25 μg/ml), and GM-CSF (50 μg/ml) respectively for 4h. The mRNA expression of endothelial activation-related markers was determined by quantitative RT-PCR. GAPDH was taken as the housekeeping gene and data was expressed as fold changes relative to control. (C) The protein expression of adhesion molecules E-selectin, VCAM1, and ICAM1 was determined by flow cytometry. (D) The protein expression of TF in HUVEC was determined by western blot. (E) The venn analysis of HUVEC stimulated with sCAR-T and TNFα. * represents p < 0.05, ** represents p < 0.01, and **** represents p < 0.0001. ns represents not significant. All data were representative of at least three independent experiments.